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Figure 5—figure supplement 2. Cells lacking Ki-67 proliferate efficiently.(A) Schematic presentation of generation of Ki-67 shRNA knockdown cell lines. (B) Top: Western blot analysis of the indicated proteins in asynchronously growing U2OS and HeLa cells stably expressing non-targeting (CTRL) or Ki-67 shRNA. Lanes separated by lines were from a single exposure of a single SDS-PAGE gel and Western blot. LC, loading control. Middle: Cell cycle distribution of these cells. Bottom: qRT-PCR analysis of the indicated mRNA levels, normalized by mRNA expression of beta-2-microglobulin (B2m). (C) Immunofluorescence of Ki-67 and EdU in asynchronous U2OS cells stably expressing control or Ki-67 shRNA, incubated with 5-ethynyl-2’-deoxyuridine (EdU) for 3 hr. Bar, 10 µm. Right: Ratio of EdU-positive cells to the total cell number for each incubation time; ns: not significant.DOI:
http://dx.doi.org/10.7554/eLife.13722.023 Image published in: Sobecki M et al. (2016) © 2016, Sobecki et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |